Detection of IL-1 alpha and IL-1 beta in the supernatants of paraformaldehyde-treated human monocytes. Evidence against a membrane form of IL-1.
The concept of a membrane form of IL-1 arose from the observation that paraformaldehyde-treated macrophages display IL-1 bioactivity. Thus far, the biochemical characterization of a membrane form of the molecule has not been reported. In a recent publication we demonstrated that murine IL-1 alpha can be detected in the supernatants of paraformaldehyde-treated macrophages. These data indicate that the phenomenon of membrane IL-1 may result from leakage of IL-1 from inadequately fixed cells. In the current report we have extended our studies toward the examination of human IL-1 alpha and IL-1 beta. IL-1 activity can be detected in the supernatants of paraformaldehyde-treated human monocytes. Although anti-IL-1 alpha, but not anti-IL-1 beta, antibodies can efficiently block the IL-1 bioactivity, both IL-1 alpha and IL-1 beta can be found by immunoprecipitation in the supernatants of the fixed monocytes. IL-1 alpha is efficiently processed to the low m.w. form, whereas IL-1 beta remains predominantly as the inactive, precursor molecule. IL-1 is not found in the supernatants of monocyte membrane preparations, demonstrating that the leakage of IL-1 is from an intracellular, rather than membrane-bound source.
Suttles
J
J
Department of Microbiology and Immunology, Wake Forest University Medical Center, Winston-Salem, NC 27103.
Carruth
L M
LM
Mizel
S B
SB
D002451:Cell Compartmentation / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects; D003593:Cytoplasm / Q000378:metabolism; D005404:Fixatives; D005557:Formaldehyde / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D007375:Interleukin-1 / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D009000:Monocytes / Q000378:metabolism* / Q000648:ultrastructure; D011108:Polymers / Q000494:pharmacology; D012995:Solubility
D005404:Fixatives; D007375:Interleukin-1; D008070:Lipopolysaccharides; D011108:Polymers; D005557:Formaldehyde; C003043:paraform